Gel-like composition and method for producing gel-like composition

Abstract

A novel gel-like composition. The present technology provides a gel-like composition containing: an enzyme having an activity; and a gelling component containing one or more gelling agents. The present technology also provides a method for producing a gel-like composition, having an enzyme adding process of adding an enzyme to a gelling component containing one or more gelling agents, in which the enzyme is added to the gelling component under temperature conditions lower than an inactivation temperature of the enzyme.

Description

Technical Field

[0001] This technology relates to gel-like compositions and methods for manufacturing gel-like compositions. Background Technology

[0002] As enzyme-containing preparations, they are sold in the form of pharmaceuticals and nutritional supplements. To date, enzyme preparations sold are generally provided in the form of tablets, capsules, granules, liquids, etc., but considering that they can be widely ingested by people of all ages and genders, there is a practical need to produce preparations in various forms.

[0003] For example, Patent Document 1 discloses an oral composition with excellent flowability, which is prepared by combining microcapsules containing water-soluble functional ingredients such as digestive enzymes in a manner that ensures good dispersibility and inhibits the dissolution of the contents.

[0004] In addition, gummy and jelly-like forms are in high demand across a wide age range, from young children to the elderly with swallowing difficulties.

[0005] For example, Patent Document 2 discloses a gummy candy that masks the unpleasant odor and taste of collagen by containing 0.5 to 7% by weight of date juice hydrolyzed by amylase.

[0006] Existing technical documents

[0007] Patent documents

[0008] Patent Document 1: Japanese Patent Application Publication No. 2011-79787

[0009] Patent Document 2: Japanese Patent Application Publication No. 2011-177036 Summary of the Invention

[0010] As mentioned above, gummy and jelly-like gel forms are in high demand, but enzyme activity may decrease during gelation. Therefore, as in Patent Document 2, even if enzymes are used in the manufacturing process of gummy candies, the enzymes will be deactivated during the manufacturing process, and gel compositions with fully residual activity are still under development.

[0011] Therefore, the main objective of this technology is to provide a novel gel-like composition.

[0012] In this technology, firstly, a gel-like composition is provided, comprising:

[0013] Enzymes, which have activity; and

[0014] The gelling component contains one or more gelling agents.

[0015] The enzyme used in the gel-like composition involved in this technology may also have an inactivation temperature higher than the gelation temperature of the gelling component.

[0016] Furthermore, the enzyme used in the gel-like composition involved in this technology may also be stable at the gelation pH of the gelling component.

[0017] The enzyme used in the gel-like composition involved in this technology is an enzyme that has the ability to be utilized in vivo.

[0018] Enzymes that can be utilized by organisms can also be enzymes that aid in digestion.

[0019] Amylase can also be an enzyme that aids digestion.

[0020] Next, a method for manufacturing a gel-like composition is provided in this technology.

[0021] It has an enzyme addition process for adding enzymes to a gelling component containing one or more gelling agents.

[0022] In the enzyme addition process, the enzyme is added to the gelling component at a temperature below the enzyme's inactivation temperature.

[0023] In the method for manufacturing the gel-like composition involved in this technology,

[0024] It may also include a gelation process that gels the gelling component that has undergone the enzyme addition process.

[0025] In the gelation process, the enzyme can also be stable. Detailed Implementation

[0026] The preferred embodiments for implementing this technology will be described below. Furthermore, the embodiments described below illustrate an example of a representative implementation of this technology, but the scope of this technology should not be interpreted narrowly as a result.

[0027] 1. Gel-like composition

[0028] The gel-like composition involved in this technology contains: an active enzyme; and a gelling component (hereinafter also referred to as the "gelling component"), which contains one or more gelling agents. The gelling component may contain gelling aids, other components, etc., as needed. The following provides a detailed description of each component.

[0029] (1) Enzyme

[0030] The enzyme contained in the gel-like composition according to this technology is characterized by its activity. That is, it is characterized in that, in the manufacturing process of the gel-like composition, for example, it does not contain an enzyme added solely for the purpose of enzymatic treatment of raw materials, but rather contains an enzyme that remains in the final product in a state of effective enzymatic activity. Furthermore, even if an enzyme is added in the manufacturing process of the gel-like composition for the purpose of enzymatic treatment of raw materials, as long as the enzyme is contained in the final product in a state of enzymatic activity, it can be used in the gel-like composition according to this technology.

[0031] To ensure that the enzyme remains in the final product in an active state, it is preferable that the enzyme is stable during the gelation step of the manufacturing method described later. Specifically, it is preferable that the enzyme is stable at the gelation temperature and gelation pH.

[0032] Typical gel compositions are liquid at high temperatures, but mostly become gels if the temperature is lower than the gelation temperature. Therefore, it is preferable that the inactivation temperature of the enzyme used is higher than the gelation temperature of the gelling component (containing the gelling agent described later and, if necessary, gelling aids, solvents, and other components). More specifically, it is preferable that the inactivation temperature of the enzyme used is at least 10°C higher than the gelation temperature of the gelling component, more preferably at least 15°C higher, even more preferably at least 20°C higher, even more preferably at least 25°C higher, and particularly preferably at least 30°C higher. By using an enzyme with an inactivation temperature within this range, temperature adjustment becomes easier when dissolving and / or dispersing the components in the liquid gelling component before gelation.

[0033] In this technology, the so-called "enzyme inactivation temperature" refers to the temperature at which the residual activity of the enzyme becomes less than 10% after the enzyme is prepared into a solution with water and placed at various temperatures for 30 minutes.

[0034] Furthermore, the inactivation pH of the enzyme used is preferably not near the gelation pH of the gelling component. More specifically, the inactivation pH of the enzyme used is preferably outside the range of ±0.1, ±0.3, ±0.5, ±1.0, and ±2.0 relative to the gelation pH of the gelling component, more preferably outside the range of ±2.5, further preferably outside the range of ±3.0, even more preferably outside the range of ±3.5, and particularly preferably outside the range of ±4.0. By using an enzyme with an inactivation pH within this range, pH adjustment during the dissolution and / or dispersion of the components in the liquid gelling component before gelation becomes easier. Additionally, in this technique, "outside the range of ±○○" means "outside the range of -○○ to +○○".

[0035] In this technique, the so-called "enzyme inactivation pH" refers to the pH at which the enzyme, after being dissolved in a buffer solution adjusted to various pH values ​​and placed at 30°C for 30 minutes, has a residual activity of less than 10%.

[0036] Provided that the function and effect of this technology are not impaired, one or more enzymes can be freely selected for use in the gel-like composition involved in this technology. In particular, the enzymes used in this technology are preferably enzymes that are bioavailable. Examples of enzymes that are bioavailable include enzymes that aid digestion and enzymes that aid metabolism. Among these, enzymes that aid digestion are preferred in this technology. Examples of enzymes that aid digestion include carbohydrate-degrading enzymes, protein-degrading enzymes, and lipid-degrading enzymes.

[0037] Examples of carbohydrate-degrading enzymes include: α-amylase, β-amylase, glucosylamylase, pullulanase, isoamylase, maltotriose hydrolase, cyclodextrin glucosyltransferase, transglucosidase, 4-α-glucantransferase, glucanase, maltase, sucrase, lactase, trehalase, cellulase, hemicellulase, pectinase, mannanase, glucosidase, hyaluronidase, and lysozyme. Examples of protein-degrading enzymes include: pepsin, trypsin, peptidase, and protease. Examples of lipid-degrading enzymes include: lipase. α-amylase and lactase will be discussed in further detail.

[0038] [α-Amylase]

[0039] α-Amylase is an enzyme that acts on starch, primarily hydrolyzing α-1,4-glucosidic bonds. Any α-amylase that can be used in this technique, provided it possesses α-amylase activity and does not impair the function and effectiveness of this technique, can also be an enzyme with other functions.

[0040] The source of α-amylase that can be used in this technology is not particularly limited, as long as it does not impair the function and effect of this technology. For example, examples of filamentous fungi include: *Aspergillus aureus*, *Aspergillus foetidus*, *Aspergillus niger*, and *Aspergillus oryzae*. Examples of actinomycetes include: *Saccharomonospora viridis*, *Streptomyces avermitilis*, *Streptomyces griseus*, *Streptomyces thermoviolaceus*, *Streptomyces violaceoruber*, and *Thermomonospora viridis*. Examples of bacteria include: *Alcaligenes latus*, *Arthrobacter*, *Bacillus amyloliquefaciens*, *Bacillus circulans*, and *Bacillus licheniformis*. The α-amylases include *Bacillus stearothermophilus*, *Bacillus subtilis*, *Cellulosimicrobium cellulans*, *Microbacterium imperiale*, *Paenibacillus alginolyticus*, and *Sulfolobus solfataricus*. Preferably, α-amylases are derived from the genus *Bacillus* or the genus *Aspergillus*, and more preferably from *Bacillus amyloliquefaciens* or from *Bacillus licheniformis*.

[0041] The content of α-amylase in the gel composition involved in this technology is not particularly limited, provided that the function and effect of this technology are not impaired. The lower limit of the α-amylase content relative to 1g of gel composition can be set as, for example, 0.004U or more, preferably 0.02U or more, more preferably 0.04U or more, 0.01U or more, 1U or more, 5U or more, and even more preferably 9U or more, 10U or more, 16U or more, 20U or more, 30U or more, 40U or more, 50U or more, 60U or more, 70U or more, or 80U or more. Furthermore, the upper limit of the α-amylase content relative to 1g of gel composition can be set, for example, below 90,000U, below 80,000U, below 70,000U, below 60,000U, below 50,000U, below 40,000U, below 30,000U, below 20,000U, below 18,000U, below 9,000U, below 8,000U, below 7,000U, below 6,000U, below 5,000U, below 400,000U. Below 0U, below 3600U, below 2000U, below 1800U, below 1000U, below 500U, below 400U, below 300U, below 200U, below 180U, below 150U, below 135U, below 120U, below 99U, below 80U, below 70U, below 60U, below 50U, below 40U, below 36U, below 27U, below 20U, below 19U, below 18U.

[0042] Furthermore, in this technique, regarding α-amylase activity, one unit (1 U) is defined as the amount of enzyme that reduces the iodine-induced color development of potato starch by 10% within one minute when treated with 1% potato starch as a substrate at pH 5.0 and 37°C. Details of the method for determining α-amylase activity in this technique will be described in the α-amylase activity determination method described in the examples below.

[0043] [Lactase]

[0044] Lactase is an enzyme that hydrolyzes lactose into galactose and glucose. Any lactase that can be used in this technique, provided it possesses lactase activity and does not impair the function and effect of this technique, can also be an enzyme with other functions.

[0045] The source of lactase that can be used in this technology is not particularly limited, as long as it does not impair the function and effectiveness of this technology. For example, examples of lactases derived from fungi include lactases from the genus *Aspergillus* such as *Aspergillus oryzae*, *Aspergillus flavus*, *Aspergillus candidus*, and *Aspergillus niger*, and lactases from the genus *Penicillium* such as *Penicillium multicolor*. Examples of lactases derived from yeast include lactases from the genus *Cryptococcus* such as *Cryptococcus terrestris* and *Cryptococcus laurentii*, lactases from the genus *Sporobolomyces* such as *Sporobolomyces singularis*, and lactases from the genus *Kluyveromyces*. Lactases from various bacteria include *Kluyveromyces* (such as *Lactis*), *Saccharomyces fragilis*, *Torula cremoris*, and *Torula utilis*. Examples of bacterial lactases include *Escherichia coli*, *Bacillus subtilis*, *Bacillus stearothermophilus*, *Bacillus circulans*, *Bacillus licheniformis*, and *Bacillus amyloliquefaciens*. Lactases from the genus *Bacillus* also exist, as do those from *Lactococcus* and *Lactobacillus bulgaricus*. Lactases from the genus *Lactobacillus* (such as *Lactobacillus*), *Streptococcus*, and *Bifidobacterium* (such as *Bifidobacterium*) are lactases derived from lactic acid bacteria.

[0046] The content of lactase in the gel composition involved in this technology is not particularly limited, provided that the function and effect of this technology are not impaired. The lower limit of the lactase content relative to 1 gram of gel composition can be set, for example, 0.05 U or more, preferably 0.25 U or more, more preferably 0.5 U or more, 1 U or more, 5 U or more, 10 U or more, 20 U or more, and even more preferably 30 U or more, 40 U or more, or 50 U or more. Furthermore, the upper limit of the lactase content relative to 1 gram of gel composition can be set as, for example, below 60,000 U, below 50,000 U, below 40,000 U, below 30,000 U, below 20,000 U, below 15,000 U, below 10,000 U, below 5,000 U, below 4,000 U, below 3,000 U, below 2,000 U, below 1,000 U, below 500 U, below 250 U, below 200 U, below 150 U, below 100 U, below 90 U, below 80 U, and below 70 U.

[0047] In this technique, lactase activity is defined as the amount of enzyme that releases 1 μmol of o-nitrophenol within 1 minute when reacting with o-nitrophenyl-β-D-galactopyranoside (ONPG) at a reaction temperature of 37°C and a reaction pH of 4.5 for 15 minutes. Details of the method for determining lactase activity in this technique will be described in the lactase activity determination method described in the examples below.

[0048] Regarding the enzymes described above, this technology is not limited to natural (wild-type) enzymes; recombinant enzymes can also be used. Commercially available enzymes or enzyme preparations can also be used. As an example of a commercially available α-amylase or α-amylase preparation, α-amylase from Bacillus amyloliquefaciens manufactured by Amano Enzyme Products Co., Ltd.

[0049] The enzymes used in this technology can be prepared from the culture medium of the microorganisms that are the source of the enzymes. Specific preparation methods include recovering enzymes from the culture medium or cells of the aforementioned microorganisms. For example, when using enzyme-secreting microorganisms, the enzymes can be separated and / or purified after recovering the cells from the culture medium by pre-filtration, centrifugation, etc., as needed. Furthermore, when using non-secreting microorganisms, the cells can be recovered from the culture medium beforehand, and then the cells can be broken up by pressure treatment, ultrasonic treatment, etc., to extract the enzymes, followed by separation and / or purification. Known protein separation and / or purification methods can be used without particular limitation for enzyme separation and / or purification, such as centrifugation, UF concentration, salting out, and various chromatographic methods using ion exchange resins. The separated and / or purified enzymes can be powdered by freeze-drying, vacuum drying, etc., and appropriate excipients and / or drying aids can also be used in this drying process. In addition, isolated and / or purified enzymes can also be made into liquids by adding appropriate additives and then filtering and sterilizing.

[0050] (2) Gelation agent

[0051] Provided that the function and effect of this technology are not impaired, one or more gelling agents that can be used in the gel-like composition involved in this technology can be freely selected. Examples of gelling agents include: pectin, carrageenan (e.g., κ-carrageenan, ι-carrageenan, λ-carrageenan), locust bean gum, guar gum, gum arabic, tamarind gum, tara gum, konjac mannan, gellan gum, xanthan gum, tragacanth gum, processed starch, gel polysaccharides, arabinoxylan, hemicellulose, cellulose and other polysaccharides; gelatin; agar; sodium alginate, etc. Among these, pectin, carrageenan (κ-carrageenan), and locust bean gum are preferred in this technology.

[0052] When using polysaccharides as gelling agents, there are no particular limitations on the source of the polysaccharides, as long as it does not impair the function and effectiveness of this technology. For example, polysaccharides from citrus peels (such as lemons, oranges, etc.), apples, beets, etc., can be used.

[0053] When using gelatin as a gelling agent, there are no particular limitations on the method of preparing the gelatin, as long as it does not impair the function and effect of this technology. For example, any substance extracted / refined from collagen-containing materials (animal skin / bone / connective tissue, etc.) is acceptable. There are also no particular limitations on the source organism of the collagen-containing material, as long as it does not impair the function and effect of this technology. For example, in addition to gelatin from the skin and bones of cattle / pigs / chickens, gelatin from aquatic organisms (freshwater / seawater) can also be used. Furthermore, substances that have undergone acid treatment, alkali treatment, etc., can also be used. In this technology, one type of gelatin with the same source and preparation method can be used, or two or more types of gelatin with different sources and preparation methods can be mixed and used.

[0054] The content of the gelling agent in the gel-like composition involved in this technology is not particularly limited, provided that the function and effect of this technology are not impaired. The lower limit of the gelling agent content can be set, for example, 0.1 w / v% or more, preferably 0.2 w / v% or more, 0.3 w / v% or more, more preferably 0.4 w / v% or more, 0.5 w / v% or more, and even more preferably 0.6 w / v% or more, 0.7 w / v% or more. Furthermore, the upper limit of the gelling agent content can be set, for example, 10 w / v% or less, preferably 9 w / v% or less, more preferably 8 w / v% or less, 7 w / v% or less, 6 w / v% or less, and even more preferably 5 w / v% or less, 4 w / v% or less, 3 w / v% or less, 2 w / v% or less, and 1 w / v% or less.

[0055] (3) Gelation aids

[0056] In this technology, a gelling aid can be used. As a gelling aid that can be used in this technology, any substance that can help gel the liquid gelling component containing the aforementioned gelling agent can be freely selected and used in one or more ways, depending on the type and amount of the gelling agent and solvent, and the type and amount of other components contained in the gelling component containing the gelling agent.

[0057] As a gelling aid, such as a pH adjuster, preferably a pH adjuster for food and beverage use, examples include organic acids such as citric acid, malic acid, tartaric acid, acetic acid, fumaric acid, and ascorbic acid; and salts such as calcium chloride.

[0058] The content of the gelling aid in the gel-like composition involved in this technology is not particularly limited, provided that the function and effect of this technology are not impaired. The lower limit of the content of the gelling aid can be set, for example, 0.1 w / v% or more, preferably 0.2 w / v% or more, 0.3 w / v% or more, more preferably 0.4 w / v% or more, 0.5 w / v% or more, and even more preferably 0.6 w / v% or more, 0.7 w / v% or more. Furthermore, the upper limit of the content of the gelling aid can be set, for example, 10 w / v% or less, preferably 9 w / v% or less, more preferably 8 w / v% or less, 7 w / v% or less, 6 w / v% or less, and even more preferably 5 w / v% or less, 4 w / v% or less, 3 w / v% or less, and 2 w / v% or less.

[0059] (4) Solvent

[0060] Provided that the function and effect of this technology are not impaired, the solvent used in the gel composition involved in this technology may be freely combined with one or more solvents that are generally used in gel compositions. Water and pH buffer solutions may be used as solvents in this technology. In addition, fruit juices and vegetable juices exemplified in the other components described below may also be used as solvents.

[0061] (5) Other ingredients

[0062] In addition, the gel-like compositions involved in this technology may freely combine one or more materials and additives used in the manufacture of general gel-like compositions. For example, they may appropriately contain fruit juice, dried fruit, vegetable juice, dried vegetables, acidulants, emulsifiers, flavorings, colorings, sweeteners, etc. Furthermore, as long as the effects and functions involved in this technology are not impaired, they may also contain inactive enzymes (e.g., enzymes used in the manufacturing process).

[0063] Fruits used in juices and dried fruits include, for example, apples, grapes, strawberries, kiwis, peaches, oranges, mangoes, and mixtures thereof. Vegetables used in vegetable juices and dried vegetables include, for example, carrots, spinach, celery, green peppers, kale, cabbage, watercress, and mixtures thereof.

[0064] (6) Final form of the gel composition

[0065] Provided that the function and effect of this technology are not impaired, the final form of the gel-like composition involved in this technology is not particularly limited. The final product can be formed directly in a gel-like form, or the final form can be formed by solidifying the gel-like composition. Specifically, examples include candies (soft candies, hard candies), gummies, chewing gum, caramel, jelly, jam, mousse, pudding, yogurt, compressed candy, tablets, etc.

[0066] 3. Method for manufacturing gel-like compositions

[0067] The method for manufacturing the gel-like composition involved in this technology includes an enzyme addition step and a gelation step. Furthermore, as needed, general gel-like composition manufacturing steps such as liquid gelation component preparation steps and recovery steps can also be performed before, after, or simultaneously with each step without compromising the effectiveness of this technology. The steps are described in detail below.

[0068] (1) Preparation process of liquid gelation components

[0069] In the process of preparing the liquid gelling component, the aforementioned gelling agent, the required gelling aid, and other components are dissolved and / or dispersed in any solvent to prepare the liquid gelling component. Furthermore, when using gelling aids and other components, they may be added during the enzyme addition step and / or gelling step described later, provided that the function and effect of this technology are not impaired. In the liquid gelling component preparation process, stirring, heating, etc., may be appropriately performed depending on the type and amount of the gelling agent, solvent, and other components used, provided that the function and effect of this technology are not impaired.

[0070] Furthermore, in the preparation process of liquid gel components, enzyme treatment of the raw materials can be performed depending on the type of solvent and other components used. After enzyme treatment, an enzyme inactivation process can also be performed appropriately.

[0071] Furthermore, the gelling agents, gelling aids, solvents, and other components used in the preparation process of the liquid gelling component are the same as those used in the above-mentioned gel composition, so they are omitted here.

[0072] (2) Enzyme addition process

[0073] The enzyme addition step is a step of adding an enzyme to a gelling component. Its characteristic is that, in the enzyme addition step, the enzyme is added to the gelling component at a temperature below the enzyme's inactivation temperature. When the aforementioned liquid gelling component preparation step is performed at a temperature below the enzyme's inactivation temperature, the liquid gelling component preparation step and the enzyme addition step can be performed simultaneously or in different orders.

[0074] As described above, in the preparation process of the liquid gelled component, if the raw material is subjected to enzyme treatment, the enzyme used in the enzyme treatment may remain in an active state. In this case, an enzyme inactivation step is not performed after enzyme treatment.

[0075] In the enzyme addition process, the specific temperature is not particularly limited as long as the enzyme can be added at a temperature below the enzyme's inactivation temperature. The enzyme addition process may be carried out at, for example, below -5°C, preferably below -10°C, more preferably below -15°C, further preferably below -20°C, and even more preferably below -25°C, relative to the inactivation temperature of the enzyme used.

[0076] In addition, during the enzyme addition process, gelling aids and other ingredients may be added as needed, as long as the function and effect of this technology are not impaired.

[0077] (3) Gelification process

[0078] The gelation process is a process of gelling the gelling component that has undergone the enzyme addition process, i.e., the liquid gelling component with added enzyme. In this technology, it is preferable that the enzyme is stable during the gelation process. By performing the gelation process under enzyme-stable conditions, it is possible to retain the enzyme in an active state in the gel-like composition.

[0079] The temperature conditions in the gelation process are lower than the inactivation temperature of the enzyme used, and can be freely set as long as the function and effect of this technology are not impaired. As for the temperature conditions in the gelation process, relative to the inactivation temperature of the enzyme used, for example, they are below -10°C, preferably below -15°C, more preferably below -20°C, further preferably below -25°C, even more preferably below -30°C, below -35°C, or below -40°C.

[0080] The pH conditions in the gelation process are not near the inactivation pH of the enzyme used, and can be freely set as long as they do not impair the function and effect of this technology. The pH conditions in the gelation process are, for example, outside the range of ±0.1, ±0.3, ±0.5, ±1.0, ±1.5, preferably outside the range of ±2.0, more preferably outside the range of ±2.5, further preferably outside the range of ±3.0, even more preferably outside the range of ±3.5, and particularly preferably outside the range of ±4.0, relative to the inactivation pH of the enzyme used.

[0081] Furthermore, during the gelation process, gelation aids and other components may be added as needed, provided that the function and effectiveness of this technology are not compromised. Specifically, examples include methods where gelation aids are added simultaneously with the gelation process, depending on the progress of the gelation. In this case, the gelation aids may be added in multiple stages.

[0082] (4) Recycling process

[0083] The recycling process is the process of recovering the gel-like composition obtained through the gelation process. The specific recycling method can be freely combined using one or more common recycling methods used in the manufacture of gel-like compositions, depending on the type of gel-like composition being produced.

[0084] Example

[0085] The present invention will now be described in more detail based on embodiments. Furthermore, the embodiments described below illustrate one example of a representative embodiment of the invention, but the scope of the invention should not be interpreted narrowly as a result.

[0086] 1. Use of raw materials / enzymes

[0087] The gelling agents and enzymes used in the examples are shown in Table 1 below.

[0088] [Table 1]

[0089]

[0090] 2. Enzyme activity assay methods

[0091] [Methods for Assaying α-Amylase Activity]

[0092] (1) Preparation of substrate solution

[0093] Approximately 1 g of potato starch was precisely measured beforehand and dried at 105 °C for 2 hours. The weight loss was then determined. To prepare a paste, the amount of potato starch corresponding to 1.000 g of the dried product was accurately measured and placed in a beaker. 20 mL of water was added, and while mixing thoroughly, 5 mL of sodium hydroxide solution (2→25) was slowly added to form a paste. Next, the paste was heated in a water bath while mixing for 3 minutes. Then, 25 mL of water was added, and after cooling, it was accurately neutralized with 2 mol / L hydrochloric acid solution. Then, 10 mL of 1 mol / L acetate-sodium acetate buffer (pH 5.0) was added, followed by the addition of water to accurately prepare 100 mL.

[0094] (2) Measurement of absorbance

[0095] Accurately measure 10 mL of the prepared substrate solution, heat at 37 ± 0.5 °C for 10 minutes, then accurately add 1 mL of the sample solution and immediately shake to mix. After accurately standing the liquid at 37 ± 0.5 °C for 10 minutes, accurately measure 1 mL of this liquid and add it to 10 mL of 0.1 mol / L hydrochloric acid solution, and immediately shake to mix. Next, accurately measure 0.5 mL of this liquid and accurately add 10 mL of 0.0002 mol / L iodine solution (NHK), shake to mix, and measure the absorbance (AT) at 660 nm using water as a control. The 0.0002 mol / L iodine solution (NHK) is prepared as follows: 10 mL of water is added to 12.7 g of iodine and 25 g of potassium iodide, and the mixture is stirred thoroughly. Water is then added to make 100 mL, which is then diluted 2500 times with water. Furthermore, 1 mL of water is accurately added instead of the sample solution, and the same procedure is performed, and the absorbance (AB) is measured.

[0096] (3) Activity calculation method

[0097] Under these conditions, the amount of enzyme that reduces the color development of potato starch caused by iodine by 10% within 1 minute is defined as 1 unit, and is calculated using the following formula.

[0098] α-Amylase activity (U / g, U / mL) = {(AB-AT) / AB} × 1 / W

[0099] AT: Absorbance of the sample solution

[0100] AB: Absorbance of blank solution

[0101] W: The amount of sample (g or mL) in 1 mL of the sample solution.

[0102] [Methods for Assaying Lactase Activity]

[0103] Measure 0.370 g of ONPG (o-nitrophenyl-β-D-galactopyranoside), add it to 0.1 mol / L acetate-sodium hydroxide buffer (pH 4.5) and dissolve to prepare 100 mL. Measure 2 mL of the ONPG solution (pH 4.5) into a 50 mL Nessler colorimetric tube, incubate at 37 ± 0.1 °C for 10 minutes, then add 0.5 mL of the sample solution and immediately vortex to mix. Incubate the liquid at 37 ± 0.1 °C for exactly 15 minutes, then add 2.5 mL of 10 g / dL sodium carbonate solution to stop the vortexing reaction. Next, add 20 mL of water and vortex to mix. The absorbance of this liquid at 420 nm was measured (A1), using water as a control. Additionally, using 0.5 mL of water instead of the sample solution as a blank, the absorbance was measured in the same manner (A2). Lactase activity was calculated using the following mathematical formula. One unit (1U) is defined as the amount of o-nitrophenol free at a rate of 1 μmol within 1 minute.

[0104] β-galactosidase activity (U / g, U / mL) = (A1-A2)×25 / ε×15×1 / w = (A1-A2)×0.7246×n

[0105] A1: Absorbance of the sample

[0106] A2: Absorbance of blank material

[0107] 25: Final liquid volume (mL)

[0108] ε: Absorption coefficient (4.60)

[0109] 15: Reaction time (minutes)

[0110] w: Enzyme amount per 0.5 mL sample solution (g, mL)

[0111] 1 / w = Dilution factor (n) per 1g or 1ml sample × 2

[0112] 3. Experimental Example

[0113] <Experimental Example 1>

[0114] (1) Preparation of gel-like composition

[0115] A gelling agent as shown in Table 2 was added to sodium acetate (pH 5.0). After dissolving the gelling agent at 50°C, the temperature was lowered to 30°C, and the enzyme shown in Table 2 was added. The temperature was then lowered to allow gelation. If gelation failed even at 20°C, a gelling aid as shown in Table 2 was added to induce gelation, thus producing a gel-like composition.

[0116] (2) Confirmation of gelation

[0117] Regarding the manufactured gel-like composition, visual inspection was conducted to confirm whether gelation had occurred at 20°C. Complete gelation was rated as ○, liquid state as ×, and semi-gelation (gel + liquid) state as △.

[0118] (3) Confirmation of enzyme activity

[0119] To confirm the enzyme activity in the manufactured gel composition, an iodine-starch reaction was performed. The gel composition was added to a substrate prepared according to the Japanese Pharmacopoeia "Test Method for Starch Dextrin Chemochemistry," and reacted at 37°C for 1 hour. Then, 2 mL of 0.0002 mol / L iodine solution (iodine.37) was added, and the color was confirmed. In the presence of residual starch, the color changed to blue-purple. If no blue-purple color change occurred, the starch was decomposed by α-amylase, indicating residual α-amylase activity. The presence of residual enzyme activity was evaluated as ○, and the absence of enzyme activity was evaluated as ×.

[0120] (4) Results

[0121] The results are shown in Table 2.

[0122] [Table 2]

[0123]

[0124] (5) Investigation

[0125] In this experimental example, a gel composition containing an active enzyme was successfully manufactured. Specifically, it was demonstrated that a gel composition containing an active enzyme can be manufactured by using an enzyme with an inactivation temperature higher than the gelation temperature of the gelling component. Furthermore, since the enzyme is stable at the gelation temperature and gelation pH of the gelling component, the enzyme activity contained in the gel composition can be sufficiently retained.

[0126] <Experimental Example 2>

[0127] (1) Preparation of gel composition A

[0128] The enzymes listed in Table 3 were added to sodium acetate (pH 6.0) at a concentration of 1% glycine and 0.5% κ-carrageenan. After dissolving at 60°C, the temperature was lowered to 30°C, and the enzymes were added. The temperature was then lowered to 20°C to allow gelation.

[0129] (2) Preparation of gel-like composition B

[0130] The mixture was added to sodium acetate (pH 6.0) at a concentration of 1% glycine, 0.5% κ-carrageenan, and 0.21% locust bean gum. After dissolving at 60°C, the temperature was lowered to 30°C, and the enzymes listed in Table 3 were added. The mixture was then cooled to 20°C to allow it to gel.

[0131] (3) Determination of residual activity

[0132] Regarding the prepared enzyme-containing gel compositions A and B, and the enzyme solutions shown in Table 3 below as Reference Examples 1 and 2, enzyme 1 (α-amylase) was stored for 1 month and enzyme 2 (lactase) was stored for 3 months at 4°C, 25°C, and 40°C. The activities of each enzyme were measured before storage and from 1 day to 90 days after storage, as recorded in Table 3 below, and the residual activity was calculated with the activity before storage as 100%.

[0133] (4) Results

[0134] The results are shown in Table 3.

[0135] [Table 3]

[0136]

[0137] (5) Investigation

[0138] As shown in Table 3, when α-amylase was used as the enzyme, the decrease in residual activity of gelled compositions A and B after storage was inhibited compared with the ungelled Reference Example 1. In particular, when stored at 40°C, the residual activity in Reference Example 1 became 0% after 1 day of storage, while the enzymes in gelled compositions A and B maintained sufficient activity.

[0139] When lactase was used as the enzyme, the enzymes in the gelled compositions A and B maintained the same activity as those in the ungelled Reference Example 2.

Claims

1. A gel-like composition comprising: Enzymes, which have activity; and The gelling component contains one or more gelling agents.

2. The gel composition according to claim 1, wherein the inactivation temperature of the enzyme is higher than the gelation temperature of the gelling component.

3. The gel composition according to claim 1, wherein the enzyme is stable at the gelation pH of the gelling component.

4. The gel composition according to claim 1, wherein the enzyme is an enzyme capable of being utilized in vivo.

5. The gel composition according to claim 4, wherein the enzyme is an enzyme that aids in digestion.

6. The gel composition according to claim 5, wherein the enzyme is an amylase.

7. A method for manufacturing a gel-like composition, It has an enzyme addition process for adding enzymes to a gelling component containing one or more gelling agents. In the enzyme addition process, the enzyme is added to the gelling component at a temperature below the enzyme's inactivation temperature.

8. The method for manufacturing the gel-like composition according to claim 7, It has a gelation process that gels the gelling component that has undergone the enzyme addition process. The enzyme is stable during the gelation process.

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